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COMPARATIVE STUDY BETWEEN ISOLATION TECHNIQUES AND PCR FORMYCOPLASMA AND UREAPLASMA DIVERSUM DETECTION IN PREPUCIAL MUCUS AND FRESH SEMEN FROM NATURAL BREEDING AND AI CENTER BOVINE BULLS.

ABSTRACT

With the objective of improving the diagnostic methods in mycoplasmosis, a study was carried out using bovine prepucial mucus and fresh semen from natural breeding and one Artificial Insemination Center with the aim to reduce the time necessary for the emission of results and to increase the detection levels, assuming bacterial isolation, on technique that depends on microbial viability, as the gold standard test. One hundred and seventy-five prepucial mucus samples and 143 fresh semen samples were studied. By PCR (Polymerase Chain Reaction) technique, a previous screening using the MGSP/GPO-1 system was tested for Mollicutes detection. Species-specific primers for Mycoplasma bovigenitalium and Ureaplasma diversumwere used subsequently. By isolation there were observed 45.1% (79/175) of positivity for Mollicutes and 66.5% (115/173) for U. diversum in prepucial mucus. Comparatively, PCR revealed 63.7% (109/171) of positive samples by MGSP/GPO-1, 42.6% (72/169) of positive samples for M. bovigenitalium and 72.9% (124/170) for U. diversum. The semen samples study presented 22.5% (32/142) of positivity for Mollicutes and 51.7% (74/143) for U. diversum, by isolation. By PCR, 24.1% of positive samples were detected (33/137) using the MGSP/GPO-1 system, 27.4% (34/124) of positive samples for M. bovigenitalium and 56.6% (73/129) for U. diversum. The McNemar test for the Mollicutes presence in prepucial mucus (p = 0.57) showed that theMGSP/GPO-1 system can have a considerable value when associated with specific detection of U. diversum, showing good sensitivity (75.2%), however lower specificity (58.9%). The statistical analysis showed that the screening by MGSP/GPO-1 system could substitute the preliminary isolation of Mycoplasma spp. in semen samples (p = 0.86).

KEY WORDS
Mycoplasma ; Ureaplasma diversum ; bulls; semen; diagnosis

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